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Practical fabrication of microfluidic platforms for live-cell microscopy

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Abstract

We describe a simple fabrication technique – targeted towards non-specialists – that allows for the production of leak-proof polydimethylsiloxane (PDMS) microfluidic devices that are compatible with live-cell microscopy. Thin PDMS base membranes were spin-coated onto a glass-bottom cell culture dish and then partially cured via microwave irradiation. PDMS chips were generated using a replica molding technique, and then sealed to the PDMS base membrane by microwave irradiation. Once a mold was generated, devices could be rapidly fabricated within hours. Fibronectin pre-treatment of the PDMS improved cell attachment. Coupling the device to programmable pumps allowed application of precise fluid flow rates through the channels. The transparency and minimal thickness of the device enabled compatibility with inverted light microscopy techniques (e.g. phase-contrast, fluorescence imaging, etc.). The key benefits of this technique are the use of standard laboratory equipment during fabrication and ease of implementation, helping to extend applications in live-cell microfluidics for scientists outside the engineering and core microdevice communities.

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Acknowledgments

We thank members of Dixon, Sims, and Holdsworth labs for their contributions, as well as members of the Skeletal Biology and Robarts Imaging Laboratories for their support. We also thank Yoshiyuki Arai for the use of his PTA ImageJ software plugin, and Mike Kovach and colleagues from Horiba-Photon Technology International for valuable advice and infrastructure support. This work was funded by the Canadian Institutes of Health Research (CIHR) grant number 126065. D. Lorusso is supported by an Ontario Graduate Scholarship and a Transdisciplinary Bone & Joint Training Award from the Collaborative Training Program in Musculoskeletal Health Research. N.M. Ochotny was supported by a Postdoctoral Fellowship Award from The Arthritis Society and by the Joint Motion Program — a CIHR Strategic Training Program in Musculoskeletal Health Research and Leadership. D. W. Holdsworth holds the Dr. Sandy Kirkley Chair in Musculoskeletal Research at The University of Western Ontario.

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Authors and Affiliations

  1. Bone and Joint Institute, The University of Western Ontario, London, Canada

    Daniel Lorusso, Noelle M. Ochotny, Stephen M. Sims, S. Jeffrey Dixon & David W. Holdsworth

  2. Imaging Research Laboratories, Robarts Research Institute, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Canada

    Daniel Lorusso, Hristo N. Nikolov, Jaques S. Milner & David W. Holdsworth

  3. Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Canada

    Daniel Lorusso, Noelle M. Ochotny, Stephen M. Sims & S. Jeffrey Dixon

  4. Department of Medical Biophysics, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Canada

    David W. Holdsworth

  5. Department of Surgery, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, Canada

    David W. Holdsworth

Authors
  1. Daniel Lorusso
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  2. Hristo N. Nikolov
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  3. Jaques S. Milner
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  4. Noelle M. Ochotny
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  5. Stephen M. Sims
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  6. S. Jeffrey Dixon
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  7. David W. Holdsworth
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Correspondence to Daniel Lorusso.

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Lorusso, D., Nikolov, H.N., Milner, J.S. et al. Practical fabrication of microfluidic platforms for live-cell microscopy. Biomed Microdevices 18, 78 (2016). https://doi.org/10.1007/s10544-016-0101-z

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  • DOI: https://doi.org/10.1007/s10544-016-0101-z

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